LOCUS       sequence-386292-        4050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 tUBQ1 terminator with GG4 GG6 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4050)
  AUTHORS   Guiziou S, Maranas CJ, Chu JC, Nemhauser JL
  TITLE     An integrase toolbox to record gene-expression during plant 
            development.
  JOURNAL   Nat Commun. 2023 Apr 3;14(1):1844. doi: 10.1038/s41467-023-37607-5.
  PUBMED    37012288
REFERENCE   2  (bases 1 to 4050)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-04-3"; volume: "14(1):1844. doi"; pages: " 
            10.1038/s41467-023-37607-5"
COMMENT     Sequence Label: L0-I15 tUBQ1 GG4-GG6
FEATURES             Location/Qualifiers
     source          1..4050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(605..624)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(606..624)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(631..648)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(631..647)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(640..662)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             785..1087
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     966..985
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1436..2230
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1436..2230
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1490..1509)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2100..2119
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2430..2439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2436..2810
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      2948..3536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3437..3456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3690..3707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3824..3845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3860..3865,3866..3883,3884..3890)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3898..3914
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3903..3925
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3922..3938
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3922..3938
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactctgtgg tctcagctta gagactctta tcaagaatcc catctcttgc ttgctttttt
       61 ttgttgtctt ccctttgata gggtttgttt ttcttgtttc agtgactttc tatgttaaaa
      121 gataatgtca gtaaaaggat ttggttttct attattctga atcgattacg gaagattctt
      181 gcttaattcc aatctataca agtatcgtga aataatgacc gtttatgtga ttaggagacg
      241 tgtttcatta ataaaatata agatcaatac attgttagta gtgataaact atgtacaaat
      301 tgtattgatt gtaaaagaaa cacaataggt tccttttttc tacaatatat tgtgacagac
      361 tctctgtttt aacgaatgaa ttaaatttgt caacgaaaaa attaaagaat gatgtgatca
      421 ctttgcacct tggttcatct tcagaggctt ctgcatgttc catgcattga ggctctctac
      481 ggtcacaggt tgtgagccgt tgttaaagac aaacaaatgc gcttgagacc tccgatcccc
      541 tgaattctgc agatatccat cacactggcg gccgctcgag catgcatcta gagggcccaa
      601 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
      661 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
      721 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt
      781 acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
      841 acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg
      901 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
      961 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     1021 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     1081 aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta
     1141 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     1201 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     1261 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     1321 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     1381 ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat
     1441 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     1501 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     1561 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     1621 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     1681 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     1741 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     1801 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     1861 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     1921 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     1981 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2041 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2101 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2161 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     2221 gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     2281 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca ataatagcac gtgaggaggg ccaccatggc caagttgacc agtgccgttc
     2461 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     2521 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     2581 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     2641 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     2701 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     2761 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctaa
     2821 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2881 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2941 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3001 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3061 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3121 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3181 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3241 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3301 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3361 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3421 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3481 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3541 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3601 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3661 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3721 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3781 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     3841 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     3901 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctattta
     3961 ggtgacgcgt tagaatactc aagctatgca tcaagcttgg taccgagctc ggatccacta
     4021 gtaacggccg ccagtgtgct ggaattcagg
//
